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Galactosylation reactions catalyzed by disaccharide phosphorylases

Chao Chen UGent (2012)
abstract
Galactosylation, as one of the main types of glycosylation reactions, is of particular importance both in vivo and in vitro. The goal of this PhD research is to explore and exploit galactosylation reactions catalyzed by disaccharide phosphorylases. On the one hand, β-galactoside phosphorylases from the novel family GH-112 were evaluated for their ability to synthesize β-galactosides. Mutational analysis of their acceptor specificity was performed, and (ga)lacto-N-biose phosphorylase was combined with lactose phosphorylase in a cascade reaction to produce bifidogenic disaccharides. On the other hand, the trehalose phosphorylase from Caldanaerobacter subterraneus (CsTP) was targeted to produce α-galactosides. More specifically, lactotrehalose was synthesized by exploiting its promiscuous activity, and the product was purified using activated carbon. Furthermore, the enzyme’s donor specificity was modified from glucose to galactose by means of iterative saturation mutagenesis, thereby producing β-galactose-1-phosphate (βGal-1P) from lactotrehalose. Based on these results, we propose the engineering of trehalose phosphorylase as a general strategy for the production of glycosyl phosphates, making use of the special features of trehalose as substrate.
Please use this url to cite or link to this publication:
author
promoter
UGent and UGent
organization
alternative title
Galactosyleringsreacties gekatalyseerd door disacharide fosforylasen
year
type
dissertation (monograph)
subject
keyword
directed evolution, sugar phosphate, donor specificity, trehalose phosphorylase, galactosylation, (ga)lacto-N-biose phosphorylase
pages
219 pages
publisher
Ghent University. Faculty of Bioscience Engineering
place of publication
Ghent, Belgium
defense location
Gent : Faculteit Bio-ingenieurswetenschappen (A0.030)
defense date
2012-10-11 16:00
ISBN
9789059895553
language
English
UGent publication?
yes
classification
D1
additional info
dissertation consists of copyrighted material
copyright statement
I have transferred the copyright for this publication to the publisher
id
3009057
handle
http://hdl.handle.net/1854/LU-3009057
date created
2012-10-08 22:20:26
date last changed
2012-10-09 09:02:30
@phdthesis{3009057,
  abstract     = {Galactosylation, as one of the main types of glycosylation reactions, is of particular importance both in vivo and in vitro. The goal of this PhD research is to explore and exploit galactosylation reactions catalyzed by disaccharide phosphorylases. On the one hand, \ensuremath{\beta}-galactoside phosphorylases from the novel family GH-112 were evaluated for their ability to synthesize \ensuremath{\beta}-galactosides. Mutational analysis of their acceptor specificity was performed, and (ga)lacto-N-biose phosphorylase was combined with lactose phosphorylase in a cascade reaction to produce bifidogenic disaccharides. On the other hand, the trehalose phosphorylase from Caldanaerobacter subterraneus (CsTP) was targeted to produce \ensuremath{\alpha}-galactosides. More specifically, lactotrehalose was synthesized by exploiting its promiscuous activity, and the product was purified using activated carbon. Furthermore, the enzyme{\textquoteright}s donor specificity was modified from glucose to galactose by means of iterative saturation mutagenesis, thereby producing \ensuremath{\beta}-galactose-1-phosphate (\ensuremath{\beta}Gal-1P) from lactotrehalose. Based on these results, we propose the engineering of trehalose phosphorylase as a general strategy for the production of glycosyl phosphates, making use of the special features of trehalose as substrate.},
  author       = {Chen, Chao},
  isbn         = {9789059895553},
  keyword      = {directed evolution,sugar phosphate,donor specificity,trehalose phosphorylase,galactosylation,(ga)lacto-N-biose phosphorylase},
  language     = {eng},
  pages        = {219},
  publisher    = {Ghent University. Faculty of Bioscience Engineering},
  school       = {Ghent University},
  title        = {Galactosylation reactions catalyzed by disaccharide phosphorylases},
  year         = {2012},
}

Chicago
Chen, Chao. 2012. “Galactosylation Reactions Catalyzed by Disaccharide Phosphorylases”. Ghent, Belgium: Ghent University. Faculty of Bioscience Engineering.
APA
Chen, Chao. (2012). Galactosylation reactions catalyzed by disaccharide phosphorylases. Ghent University. Faculty of Bioscience Engineering, Ghent, Belgium.
Vancouver
1.
Chen C. Galactosylation reactions catalyzed by disaccharide phosphorylases. [Ghent, Belgium]: Ghent University. Faculty of Bioscience Engineering; 2012.
MLA
Chen, Chao. “Galactosylation Reactions Catalyzed by Disaccharide Phosphorylases.” 2012 : n. pag. Print.